CELL IMAGING AND HISTOLOGY CORE 1. MAIN OBJECTIVES AND NEW DIRECTIONS Under the NIH Neuroscience Blueprint Core Center Grant, the Cell Imaging/Histology Core will be built as an expansion of an existing Cancer Center core at the Burnham Institute for Medical Research (BIMR). As described elsewhere in this grant application, historically, Cancer Center cores have not allowed access to neuroscientists on the La Jolla Torrey Pines Mesa. This Neuroscience Center Grant will provide equal access and foster novel Neuroscience-specific imaging tools and histology-related applications for the Neuroscience community. The Core will also advance the molecular and cellular understanding of neural development, function and pathology, and serve as an initiator for collaborations in the Neuroscience community throughout San Diego. Current neuroscience research widely employs genetic manipulations to address the workings of specific molecules in model organisms and neural cultures. The observation of the molecular changes implemented in these systems critically depends on histological/immunohistological and advanced molecular imaging techniques. The mission of the Neuro-lmaging and Histology Core is to provide state-of-the-art imaging and histological tools to the Neuroscience community on the Torrey Pines Mesa. Currently, Neuroscientists at the Salk Institute and BIMR have only limited access to expertise and instrumentation in neuronal imaging and histology of the nervous system. The Salk Institute operates a standard single photon confocal microscope (Leica TCS SP2 AOBS) on a shared basis that is suited for molecular imaging and extensively used by Salk researchers. However, the capabilities of this instrument are limited, in particular when it comes to imaging of cellular or molecular events in thick tissue preparations, or to live tissue preparations. The Burnham Institute's existing Cell Imaging and Histology Facility has available within its well-equipped Core, the instrumentation, expertise and capacity for multiphoton, confocal imaging that will accommodate this void. Neither the existing Salk nor the BIMR facilities offer services geared specifically to Neuroscience applications. The current BIMR service for histology is well versed for the preparation and analysis of tumors and large organs, but cannot accommodate tasks geared towards analyzing specific structures within the complex architecture of the nervous system. The goal for this Neuroscience Blueprint Initiative is to overcome these limitations and establish an open access Core Facility for neuroscientists on the Torrey Pines Mesa. The Burnham Institute's Cancer Center Cell Imaging Facility has experienced significant expansion during the past year and invested in three new confocal microscopes as part of its institutional commitment to expand use and capabilities. With these new instruments, the imaging capabilities at BIMR are significantly improved and additional imaging capacity is now open. Recent techniques in molecular imaging have experienced significant advancement with the development by Dr. Roger Tsien of UCSD of fluorescent dyes that allow the tracing of molecules within living cell. Dr. Tsien will bring the imaging tools he has developed to this Neuroscience Blueprint Core and participate in "spying" on the molecules of the nervous system as the Co-Director. Thus, users will gain easy access to the tools developed by Dr. Tsien through this Core. For the histology section of the Core, specialized on-site expertise in the anatomy and histology of the nervous system will be added to the existing Cancer Center BIMR Histology facility. The Core will accommodate the preparation of brain sections to view specified anatomical structures in the developing and adult nervous system. Moreover, the Core will operate a state-of-the-art laser microdissection system that will allow excising specified cells or nuclei for microarray and proteomic profiling. Thus, the Imaging/Histology Core will interphase with the Gene Analysis and the Proteomics Cores of this application. Electron microscopy (EM) is not included in this Imaging/Histology Core as Neuroscience researchers will have access to excellent ultrastructural analyses of the nervous system through the Neuropathology Core in this grant, which is directed by Dr. Eliezer Masliah (see Neuropathology Core Description in this application). Through the Neuropathology Core, Neuroscientists on the Torrey Pines Mesa also have priority access to the National Center for Microscopy and Imaging Research (NCMIR) Facility directed by Dr. Mark Ellisman of UCSD. The NCMIR Facility supports scanning-, transmitted-, and high voltage electron microscopy. The mission for the Cell Imaging and Histology Core is to extend molecular and cellular imaging capabilities and histology services to the Neuroscience community on the Torrey Pines Mesa. The main Core objectives are: 1) Provide high-resolution microscopic techniques in combination with molecular probes to obtain information about the location, movement, and activities of molecules within cells; 2) Provide histological services to reveal the anatomical structures and cellular components of the developing and adult nervous system, and for the laser dissection of cell populations for gene array and proteome analysis. 3) Offer consultation, train users and assist with the analysis of the data generated within the Core. 4) Foster collaborations between neuroscientists through information exchange and interphase work with the other Cores of this Blueprint Initiative.